The presence of non-coding RNAs, such as microRNAs (miRNAs), in mitochondria has been reported by several studies. The biological roles and functions of these mitochondrial miRNAs ("mitomiRs") have not been sufficiently characterized, but the mitochondrial localization of miRNAs has recently gained significance due to modified mitomiR-populations in certain states of diseases. Here, we describe the isolation and analysis of mitochondrial RNAs from rat liver tissue and HepG2 cells. The principle of the analysis is to prepare mitochondria by differential centrifugation. Cytosolic RNA contamination is eliminated by RNase A treatment followed by percoll gradient-purification and RNA-extraction. Small RNA content is verified by capillary electrophoresis. Mitochondrial miRNAs are detected by Q-PCR following synthesis of cDNA. After Q-PCR based mitomiR-profiling, the Normfinder algorithm is applied to identify suitable reference miRNAs to use as normalizers for mitochondrial input and data analysis. The described procedure depicts a simple way of isolating and quantifying mitomiRs in tissue and cell culture samples.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/978-1-0716-4264-1_14 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Who is coming in? Evaluation of physician performance within multi-physician emergency departments.
Am J Emerg Med
January 2025
Department of Emergency Medicine, Yale University School of Medicine, New Haven, CT, USA; Center for Outcomes Research and Evaluation, Yale University, New Haven, CT, USA.
Background: This study aimed to examine how physician performance metrics are affected by the speed of other attendings (co-attendings) concurrently staffing the ED.
Methods: A retrospective study was conducted using patient data from two EDs between January-2018 and February-2020. Machine learning was used to predict patient length of stay (LOS) conditional on being assigned a physician of average speed, using patient- and departmental-level variables.
Human fallopian tube organoids provide a favourable environment for sperm motility.
Hum Reprod
January 2025
IRSD, Université de Toulouse, INSERM, INRAE, ENVT, Univ Toulouse III-Paul Sabatier (UPS), Toulouse, France.
Study Question: Does a human fallopian tube (HFT) organoid model offer a favourable apical environment for human sperm survival and motility?
Summary Answer: After differentiation, the apical compartment of a new HFT organoid model provides a favourable environment for sperm motility, which is better than commercial media.
What Is Known Already: HFTs are the site of major events that are crucial for achieving an ongoing pregnancy, such as gamete survival and competence, fertilization steps, and preimplantation embryo development. In order to better understand the tubal physiology and tubal factors involved in these reproductive functions, and to improve still suboptimal in vitro conditions for gamete preparation and embryo culture during IVF, we sought to develop an HFT organoid model from isolated adult stem cells to allow spermatozoa co-culture in the apical compartment.
Collapsing glomerulopathy associated with parvovirus B19 and systemic lupus erythematosus in a patient with APOL1 high-risk variant for nephropathy.
J Bras Nefrol
January 2025
Universidade Federal de São Paulo, Departamento de Medicina, São Paulo, SP, Brazil.
Collapsing glomerulopathy (CG) has a severe course typically associated with viral infections, especially HIV and parvovirus B19, systemic lupus erythematosus (SLE), among other etiologies. A 35-year-old woman with recent use of a JAK inhibitor due to rheumatoid arthritis presented with a 2-week history of fever, cervical adenopathy, and facial erythema. After admission, anemia, hypoalbuminemia, proteinuria, and severe acute kidney injury were noted.
View Article and Find Full Text PDFPheno-Morphological Screening and Acoustic Sorting of 3D Multicellular Aggregates Using Drop Millifluidics.
Adv Sci (Weinh)
January 2025
CNRS, Univ. Bordeaux, CRPP, UMR 5031, Pessac, F-33600, France.
Three-dimensional multicellular aggregates (MCAs) like organoids and spheroids have become essential tools to study the biological mechanisms involved in the progression of diseases. In cancer research, they are now widely used as in vitro models for drug testing. However, their analysis still relies on tedious manual procedures, which hinders their routine use in large-scale biological assays.
View Article and Find Full Text PDFTranscriptomic HIV-1 reservoir profiling reveals a role for mitochondrial functionality in HIV-1 latency.
PLoS Pathog
January 2025
Department of Experimental Immunology, Amsterdam UMC Location University of Amsterdam, Amsterdam, Netherlands.
Identifying cellular and molecular mechanisms maintaining HIV-1 latency in the viral reservoir is crucial for devising effective cure strategies. Here we developed an innovative flow cytometry-fluorescent in situ hybridization (flow-FISH) approach for direct ex vivo reservoir detection without the need for reactivation using a combination of probes detecting abortive and elongated HIV-1 transcripts. Our flow-FISH assay distinguished between HIV-1-infected CD4+ T cells expressing abortive or elongated HIV-1 transcripts in PBMC from untreated and ART-treated PWH from the Amsterdam Cohort Studies.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!